Anti-ESAM antibody (ab74777)

Overview

  • Product name
  • Description
    Rabbit polyclonal to ESAM
  • Tested applications
    Suitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Cynomolgus monkey
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human ESAM.

    (Peptide available as ab101730.)

  • Positive control
    • This antibody gave a positive signal in the following lysates: Human Heart tissue; Human Liver tissue; Human Kidney tissue; Human Lung tissue; HepG2 whole cell; HEK293 whole cell.

Properties

Applications

Our Abpromise guarantee covers the use of ab74777 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 41 kDa).

Target

  • Function
    Can mediate aggregation most likely through a homophilic molecular interaction.
  • Tissue specificity
    Highly expressed in endothelial cells.
  • Sequence similarities
    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Cellular localization
    Cell junction > adherens junction. Cell junction > tight junction. Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • 2310008D05Rik antibody
    • Endothelial cell adhesion molecule antibody
    • Endothelial cell selective adhesion molecule antibody
    • Endothelial cell-selective adhesion molecule antibody
    • Esam antibody
    • ESAM_HUMAN antibody
    • HUEL (C4orf1) interacting protein antibody
    • LP4791 protein antibody
    • W117m antibody
    see all

Images

  • All lanes : Anti-ESAM antibody (ab74777) at 1 µg/ml

    Lane 1 : Human heart tissue lysate - total protein (ab29431)
    Lane 2 : Human liver tissue lysate - total protein (ab29889)
    Lane 3 : Human kidney tissue lysate - total protein (ab30203)
    Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 6 : Lung (Human) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Annexin A2 overexpression lysate (ab94080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 41 kDa
    Observed band size : 37,41 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 100 kDa,15 kDa,250 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes
  • ICC/IF image of ab74777 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab74777, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2 cells at 5µg/ml, and in 100% methanol fixed (5 min) HepG2 and MCF7 cells at 5µg/ml.

References

ab74777 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colon tumour)
Specification
Colon tumour
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH6.0
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
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Submitted Nov 08 2011

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